Three-Dimensional Spatio-Temporal Control of Lipid Nanoparticle Manufacturing for Improved Nucleic Acid Delivery

PROJECT SUMMARY The COVID-19 pandemic has established the importance of nucleic acid-based lipid nanoparticles (LNPs) for the future of global health. The benefits of lipid nanoparticles are multifaceted as they protect sensitive pharmaceutical payloads from enzymatic degradation and allow for the modification of solubility, release kinetics, and bioavailability. While chemical formulation of LNPs has been widely explored, the effects of manufacturing—specifically microfluidics processing—are not currently well-understood. This knowledge gap presents challenges in the production of intricate nanoparticle structures, which require specialized microfluidic systems that produce well-defined and reproducible flow configurations. The proposed research focuses on developing 3D-printed channel architectures to precisely control LNP structure and properties to enhance transfection efficiency without modifying their chemical composition. Aim 1 involves designing, simulating, and testing various 3D channel architectures to manipulate flow conditions and tailor LNP properties. Aim 2 focuses on structural determination via SAXS and CryoTEM, and assesses the impact of LNP structures on mRNA transfection efficiency through in vitro transfection studies. Aim 3 will demonstrate the therapeutic- and disease-agnostic design workflow by robustly encapsulating siRNA and pDNA. This project aims to overcome the limitations of current LNP manufacturing methods which are constrained by fixed geometries and limited control over LNP assembly processes. Development of our enabling technology will offer an additional process parameter - channel architecture, for tuning LNP properties and structure in a rapid and customizable manner that is broadly applicable.